Simple Microfluidics Research Articles

Patterned delivery and expression of gene constructs into zebrafish embryos using microfabricated interfaces

We demonstrate a method which uses simple microfabrication and microfluidics to produce custom, shaped electroporators for the patterned delivery of foreign molecules into developing embryos. We show how these electroporators can be used to 'draw' two-dimensional patterns of tracer molecules, DNA and mRNA into the yolk and cells of zebrafish embryos (Danio rerio) at different stages of development. We demonstrate the successful delivery of patterns of Trypan Blue (normal dye), Texas Red (fluorescent dye), GFP-expressing DNA plasmids and GFP expressing mRNA constructs into both chorionated and dechorionated embryos. Both DNA and mRNA were expressed in the desired patterns subsequent to delivery. Square pulses of 10-20 V (0.20-0.40 kV/cm), 50-100 ms width were sufficient to create transient pores and introduce compounds from the late blastula period (3 hpf) to early pharyngula period (24 hpf) embryos. Using 24 hpf dechorionated embryos, we achieved a high survival of 91.3% and 89%, and a delivery efficiency of 38% and 50% for GFP-DNA and GFP-mRNA respectively. Lastly, we demonstrate the simultaneous delivery of different compounds into the developing embryo.

204 POSTER Discovery and validation of a promising new target for therapeutic monoclonal antibodies: a type II transmembrane serine protease overexpressed in human ovarian and pancreatic cancers

When a sessile droplet of a complex fluid dries, a stain forms on the solid surface. The structure and pattern of the stain can be used to detect the presence of a specific chemical compound in the sessile droplet. In the present work, we investigate what parameters of the stain or its formation can be used to characterize the specific interaction between an aqueous dispersion of beads and its receptor immobilized on the surface. We use the biotin-streptavidin system as an experimental model. Clear dissimilarities were observed in the drying sequences on streptavidin-coated substrates of droplets of aqueous solutions containing biotin-coated or streptavidin-coated beads. Fluorescent beads are used in order to visualize the fluid flow field. We show differences in the distribution of the particles on the surface depending on biomolecular interactions between beads and the solid surface. A mechanistic model is proposed to explain the different patterns obtained during drying. The model describes that the beads are left behind the receding wetting line rather than pulled towards the drop center if the biological binding force is comparable to the surface tension of the receding wetting line. Other forces such as the viscous drag, van der Waals forces, and solid–solid friction forces are found negligible. Simple microfluidics experiments are performed to further illustrate the difference in behavior where is adhesion or friction are present between the bead and substrate due to the biological force. The results of the model are in agreement with the experimental observations which provide insight and design capabilities. A better understanding of the effects of the droplet–surface interaction on the drying mechanism is a crucial first step before the identification of drying patterns can be promisingly applied to areas such as immunology and biomarker detection.

Local Heating of Discrete Droplets Using Magnetic Porous Silicon-Based Photonic Crystals

This paper describes a method for local heating of discrete microliter-scale liquid droplets. The droplets are covered with magnetic porous Si microparticles, and heating is achieved by application of an external alternating electromagnetic field. The magnetic porous Si microparticles consist of two layers. The top layer contains a photonic code and it is hydrophobic, with surface-grafted dodecyl moieties. The bottom layer consists of a hydrophilic silicon oxide host layer that is infused with Fe3O4 nanoparticles. The amphiphilic microparticles spontaneously align at the interface of a water droplet immersed in mineral oil, allowing manipulation of the droplets by application of a magnetic field. Application of an oscillating magnetic field (338 kHz, 18 A rms current in a coil surrounding the experiment) generates heat in the superparamagnetic particles that can raise the temperature of the enclosed water droplet to >80 degrees C within 5 min. A simple microfluidics application is demonstrated: combining complementary DNA strands contained in separate droplets and then thermally inducing dehybridization of the conjugate. The complementary oligonucleotides were conjugated with the cyanine dye fluorophores Cy3 and Cy5 to quantify the melting/rebinding reaction by fluorescence resonance energy transfer (FRET). The magnetic porous Si microparticles were prepared as photonic crystals, containing spectral codes that allowed the identification of the droplets by reflectivity spectroscopy. The technique demonstrates the feasibility of tagging, manipulating, and heating small volumes of liquids without the use of conventional microfluidic channel and heating systems.

Manipulation of liquid droplets using amphiphilic, magnetic one-dimensional photonic crystal chaperones

The controlled manipulation of small volumes of liquids is a challenging problem in microfluidics, and it is a key requirement for many high-throughput analyses and microassays. One-dimensional photonic crystals made from porous silicon have been constructed with amphiphilic properties. When prepared in the form of micrometre-sized particles and placed in a two-phase liquid such as dichloromethane/water, these materials will accumulate and spontaneously align at the interface. Here we show that superparamagnetic nanoparticles of Fe(3)O(4) can be incorporated into the porous nanostructure, allowing the materials to chaperone microlitre-scale liquid droplets when an external magnetic field is applied. The optical reflectivity spectrum of the photonic crystal displays a peak that serves to identify the droplet. Two simple microfluidics applications are demonstrated: filling and draining a chaperoned droplet, and combining two different droplets to perform a chemical reaction. The method provides a general means for manipulating and monitoring small volumes of liquids without the use of pumps, valves or a microfluidic container.

Perfectly monodisperse microbubbling by capillary flow focusing.

Here we report a simple microfluidics phenomenon which allows the efficient mass production of micron size gas bubbles with a perfectly monodisperse and controllable diameter. It resorts on a self-excited breakup phenomenon (which locks at a certain frequency) of a short gas microligament coflowing in a focused liquid stream. In this work, we describe the physics of the phenomenon and obtain closed expressions for the bubble diameter as a function of the liquid and gas properties, geometry, and flow parameters, from a large set of experimental results.

Flow of Eringen fluid (simple microfluid) through an artery with mild stenosis

This paper presents an analysis for blood flow through a stenosed artery considering the localized effects of the stenosis. The blood vessel is modelled as a rigid tube and the particulate nature of blood is accounted through an Eringen fluid (simple microfluid) model. Further a peripheral layer of Newtonian fluid has been considered in the model to account for the cell free layer. It is observed that the resistance to the flow as well as the wall shear stress increase as the height of the stenosis increases. However, the trend is reversed as the peripheral layer thickness increases.

Dispersion in simple microfluid flows

We have discussed here dispersion of a solute matter in a simple microfluid flowing through channels and pipes. The effect of homogeneous first order chemical reaction rate has also been considered in the analysis. Using Taylor's limiting condition, equivalent dispersion coefficient has been determined and the effects of various parameters have been studied. Moreover, as the various viscosity coefficients for simple microfluid are known to depend on the volume concentration of suspended particles, its effect on the equivalent dispersion coefficient has also been studied. It is found that for given simple microfluid parameters the equivalent dispersion coefficient decreases as the chemical reaction rate constant increases, however it increases as the volume concentration of suspended particles increases.

The theory on homogeneous micro-fluid mixtures

This paper deals with the construction x of a generalized- diffusive theory of simple micro-fluid mixtures composed of a reactive constituents in the relative motion to one another and treated as a single simple micro-fluid. The basic laws of balance are given by postulating the balance law of energy together with invariant requirement under superposed rigid body motions or the mixture as a whole. The linear or nonlinear constitutive equations and full thermodynamic restrictions are given.

On nonlocal microfluid mechanics

The simple microfluid theory of Eringen [1] is generalized to include nonlocal effects. The balance laws, jump conditions, and constitutive equations are obtained. The nonlocal intermolecular forces, energy and entropy are accounted for by nonlocal field residuals and functional constitutive equations of space type. The second law of thermodynamics is used to obtain specific forms of these residuals, constitutive equations and restrictions to be imposed. The linear theory is developed fully for the micromorphic and micropolar nonlocal fluids. The nonlocal effects are shown to include surface tension and surface stresses. Passage is made to material gradient theories.

Micromorphic description of turbulent channel flow

The simple microfluid theory of Eringen is employed to determine the velocity profile and microgyrations in a steady flow of viscous fluids between two parallel walls. The shearing stress, microrotations, microinertia and Reynold stresses are determined. The results are compared with the experimental work of Laufer.

Theory of thermomicrofluids

The simple microfluid theory of Eringen [1] is extended to include the heat conduction and heat dissipation effects. The exact nonlinear theory is presented and restricted by the axioms of constitution and the second law of thermodynamics. The constitutive equations are linearized for dissipative and thermal effects. The complete field equations and accompanying jump conditions are given. Several internally constrained fluids, e.g., micropolar, inertia rateless fluids, are obtained. Boundary and initial conditions are discussed.